Mabs against ICAM-1/CD54 in rhesus monkey

From: Sukwan Handali (shandau@mailhost.tcs.tulane.edu)
Date: Mon Jun 12 2000 - 10:50:26 EST


To whom it may concern!

I am a student at Dept. Tropical Medicine, Tulane University. I am detecting
ICAM-1/CD54 from rhesus monkey brain endothelial cells. As my positive
control, human brain endothelial cells. I have detected ICAM-1 in human
brain endothelial cells (constitutive and upregulated by TNF), but I failed
to detect ICAM-1 in rhesus monkeys.

The MAbs that I have used: 84H10 (Immunotech) and LB-2 clone (BD). LB-2
clone according to BD will cross-react to ICAM-1 of rhesus monkey (based on
a web-site at Univ. Florida).

So, my questions:

1.    what should I do so that I could detect ICAM-1 from rhesus monkey?
2.    is trypsinization will destroy ICAM1 from rhesus monkeys? (my positive
control is trypsinized and works well)
3.    I have tried to detach my cells with EDTA 1 mM in PBS and it did not
work. Will Sigma Cell Detachment Solution will work? Or, if somebody knows
how to detach this kind of sticky cells (without scrapping and trypsin) and
still preserve ICAM-1?
4.    is there somebody who has work with ICAM-1 in rhesus monkey? I wonder
if the researcher(s) who put the information on U Florida website could
share the procedure!

Thanks.


Sukwan Handali



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